EP0684763B1 - Method and apparatus for treating a body fluid - Google Patents

Method and apparatus for treating a body fluid Download PDF

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Publication number
EP0684763B1
EP0684763B1 EP95904869A EP95904869A EP0684763B1 EP 0684763 B1 EP0684763 B1 EP 0684763B1 EP 95904869 A EP95904869 A EP 95904869A EP 95904869 A EP95904869 A EP 95904869A EP 0684763 B1 EP0684763 B1 EP 0684763B1
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EP
European Patent Office
Prior art keywords
viral
agent
blood
body fluid
column
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Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Expired - Lifetime
Application number
EP95904869A
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German (de)
French (fr)
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EP0684763A4 (en
EP0684763A1 (en
Inventor
John T. Foley
John Chapman
Ludwig Wolf, Jr.
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Baxter International Inc
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Baxter International Inc
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Publication date
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Publication of EP0684763A4 publication Critical patent/EP0684763A4/en
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Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
    • A61M1/00—Suction or pumping devices for medical purposes; Devices for carrying-off, for treatment of, or for carrying-over, body-liquids; Drainage systems
    • A61M1/02—Blood transfusion apparatus
    • A61M1/0209—Multiple bag systems for separating or storing blood components
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L2/00—Methods or apparatus for disinfecting or sterilising materials or objects other than foodstuffs or contact lenses; Accessories therefor
    • A61L2/0005—Methods or apparatus for disinfecting or sterilising materials or objects other than foodstuffs or contact lenses; Accessories therefor for pharmaceuticals, biologicals or living parts
    • A61L2/0082—Methods or apparatus for disinfecting or sterilising materials or objects other than foodstuffs or contact lenses; Accessories therefor for pharmaceuticals, biologicals or living parts using chemical substances
    • A61L2/0088—Liquid substances
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
    • A61M1/00—Suction or pumping devices for medical purposes; Devices for carrying-off, for treatment of, or for carrying-over, body-liquids; Drainage systems
    • A61M1/02—Blood transfusion apparatus
    • A61M1/0209—Multiple bag systems for separating or storing blood components
    • A61M1/0218—Multiple bag systems for separating or storing blood components with filters
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
    • A61M1/00—Suction or pumping devices for medical purposes; Devices for carrying-off, for treatment of, or for carrying-over, body-liquids; Drainage systems
    • A61M1/36—Other treatment of blood in a by-pass of the natural circulatory system, e.g. temperature adaptation, irradiation ; Extra-corporeal blood circuits
    • A61M1/3681—Other treatment of blood in a by-pass of the natural circulatory system, e.g. temperature adaptation, irradiation ; Extra-corporeal blood circuits by irradiation
    • A61M1/3683—Other treatment of blood in a by-pass of the natural circulatory system, e.g. temperature adaptation, irradiation ; Extra-corporeal blood circuits by irradiation using photoactive agents
    • A61M1/3686—Other treatment of blood in a by-pass of the natural circulatory system, e.g. temperature adaptation, irradiation ; Extra-corporeal blood circuits by irradiation using photoactive agents by removing photoactive agents after irradiation
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
    • A61M1/00—Suction or pumping devices for medical purposes; Devices for carrying-off, for treatment of, or for carrying-over, body-liquids; Drainage systems
    • A61M1/36—Other treatment of blood in a by-pass of the natural circulatory system, e.g. temperature adaptation, irradiation ; Extra-corporeal blood circuits
    • A61M1/3687—Chemical treatment

Definitions

  • the present invention relates generally to the collection and therapeutic use of body fluids. More specifically, the present invention relates to methods reducing or eliminating potential viral contaminants and other pathogens in body fluids, such as blood.
  • body fluids especially blood components, such as red blood cells, platelets, plasma, and bone marrow
  • therapies provide treatments, many of which are life saving, and cannot otherwise be provided, due to the transmission of infectious diseases, there may be potential risks to such therapies.
  • blood can carry infectious agents, such as hepatitis virus, human immune deficiency virus (an etiological agent for AIDS), cytomegalovirus, Epstein Barr virus, and herpes virus.
  • infectious agents such as hepatitis virus, human immune deficiency virus (an etiological agent for AIDS), cytomegalovirus, Epstein Barr virus, and herpes virus.
  • screening methods exist to identify blood that may include such viruses, current screening methods do not assure that every blood unit that contains such a virus is identified.
  • a contaminated blood component will only exhibit a positive test if it includes antibodies for the virus, e.g., anti-HIV.
  • an individual may not generate antibodies immediately upon infection. Rather, there is a window period that extends from the initial infection of the patient with a virus to the generation of antibodies. When an individual is in this window period, diagnostic tests that are based on antibodies will not identify the individual, or the blood unit, as being infected. But, even though the antibodies are not present, the blood unit can still transmit an infection.
  • this window period can extend from approximately six weeks up to 48 months.
  • an individual who has been infected with HIV and accordingly, whose blood will transmit same, will register a negative antibody response.
  • screening methods will not identify as contaminated a blood unit from an individual who is infected with HIV, but who has not generated anti-HIV.
  • photoactive agents that have antiviral action have been explored.
  • These photoactive agents are generally agents that upon activation with light will inactivate or destroy pathogens, e.g., a virus that may be present.
  • Such photoactive agents include: psoralens; porphyrins; phthalocyanines; and dyes, such as methylene blue. See, for example, U.S. Patent Nos.: 4,748,120; 4,878,891; 5,120,649; and German Patent Application No. DE 39 30 510 A1 (Mohr).
  • EP-A-0124363 and WO-A-91/03933 disclose treatment of body fluids with a photoactive agent and irradiating the fluids to inactivate viral contaminants.
  • EP-A-0124363 discloses removal of the agent and photoproducts by dialysis and WO-A-91/03933 discloses removal of the agent by dialysis, or gel filtration and specifically by use of Bio Beads.
  • methylene blue is a photoactive agent that has been shown to have efficacy in inactivating viral contamination in plasma.
  • methylene blue has been, through exhaustive testing, shown to have no toxicity concerns, upon photoactivation of methylene blue, photoproducts are generated.
  • Azure A and B are generated upon photoactivation of methylene blue. The in vivo effect of these products has not been as well studied as methylene blue in patients and therefore they raise regulatory issues and in vivo concerns.
  • a viral inactivation agent to a body fluid is added a viral inactivation agent.
  • the resultant product is then passed through a container, e.g., column including a material having an affinity for the viral inactivating agent. This allows the column to remove excess viral inactivating agent. Additionally, other products, e.g. photoproducts, that may be generated upon addition of the viral inactivation agent or any activation thereof are also eliminated.
  • the body fluid can then be infused into a patient without regulatory or toxicity concerns.
  • the material includes charcoal.
  • the column is an ion exchange column.
  • the blood product includes platelets and the viral inactivating agent is a psoralen.
  • the blood product includes plasma and the viral inactivating agent includes methylene blue.
  • pathogens from blood or its components are removed before they are infused into a patient and any post thaw photoactivation of excess photoactivated agents is prevented.
  • Another advantage of the present invention is that it allows normal plasma color for treated plasma.
  • FIG 1 illustrates, schematically, an embodiment of apparatus for carrying out the present invention.
  • the present invention provides a method for use in treating a body fluid, such as blood, to reduce or eliminate viral contaminations that may be present therein. It is believed that the present invention can be used with a variety of photoactive viral inactivating agents, such as psoralens, porphyrins and dyes, such as methylene blue, phthalocyanines, phenothiazines, hypericin, and other compounds that are activated by light.
  • photoactive viral inactivating agents such as psoralens, porphyrins and dyes, such as methylene blue, phthalocyanines, phenothiazines, hypericin, and other compounds that are activated by light.
  • a photoactive viral inactivating agent would be added to a body fluid, such as blood prior to the blood being infused into a patient.
  • the resultant blood product including the photoactive agent then would be activated by light of a suitable wavelength.
  • a container 10 including a blood component 11.
  • the blood component has added thereto a viral inactivation agent 13.
  • the blood component can be added to the container 10 including the viral inactivation agent.
  • the viral inactivation agent for example, methylene blue can be added to the plasma component.
  • whole blood can be treated with a viral inactivation process.
  • a separate container is not required and the viral inactivation agent can be added to the container in whcih the component is stored.
  • the container 10 will include a fluid line 12 that will be coupled to a column 14.
  • column refers broadly to a chamber or device that includes material that will remove specific compounds or entities. Accordingly, column includes cartridges, containers, and other means for housing such material.
  • the column will include an inlet 16 allowing product to flow into an interior 18 defined by the housing 20.
  • porous plates are located at each end 26 and 28, respectively, of the interior 18 of the housing 20. The porous plates allow the body fluid to flow through an affinity matrix located therein. The resultant product then flows out of the column 14 through the outlet 34.
  • the resultant product flows through fluid line 12 and into the affinity column 14.
  • the affinity column 14 will remove excess viral inactivation agents, as well as photoproducts. For example, with respect to methylene blue, excess methylene blue will be removed, as well as Azure A and B.
  • the resultant blood product will then flow through fluid line 36 to a container 38.
  • the blood can be stored in the container 38 and then infused into a patient.
  • breakable cannulas as are known in the art, can be provided. Of course, other means for allowing selective flow through the fluid line 12 can be provided.
  • the column 14 is a separate and distinct component from the container 10, a unitary structure can be provided.
  • the column can be integral with the container or coupled thereto as an outlet port of the container.
  • the material used for the matrix in the affinity column 14 comprises charcoal or an ion exchange resin.
  • AMT 4'-aminomethyl-4,5',8-trimethyl psoralen
  • a second charcoal column was tested at a flow rate of about 5 mL/min. This column removed "100%" of the AMT as measured by HPLC. Platelet loss was 14%. Total protein increased by 14%. The platelet morphology score was unchanged by the column (200).
  • the ion exchange column clearly removed significant amounts of AMT at low flow rate, but not as much as the charcoal. This column did not appear to remove any plasma protein and platelet loss was higher than with the charcoal. The platelets did not appear to be effected by the ion exchange resin.

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  • Health & Medical Sciences (AREA)
  • Heart & Thoracic Surgery (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Public Health (AREA)
  • Biomedical Technology (AREA)
  • Engineering & Computer Science (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Vascular Medicine (AREA)
  • Veterinary Medicine (AREA)
  • Anesthesiology (AREA)
  • Hematology (AREA)
  • Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Medicinal Chemistry (AREA)
  • Molecular Biology (AREA)
  • Epidemiology (AREA)
  • Cardiology (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)
  • External Artificial Organs (AREA)
  • Devices And Processes Conducted In The Presence Of Fluids And Solid Particles (AREA)
  • Electrical Discharge Machining, Electrochemical Machining, And Combined Machining (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)

Abstract

A method for treating a body fluid to at least substantially inactivate viral contaminants that may be present therein is provided. The method includes the steps of: providing a body fluid; adding to the body fluid a viral inactivating agent to create a resultant product; and passing the resultant product through a column including material having an affinity for the viral inactivating agent.

Description

BACKGROUND OF THE INVENTION
The present invention relates generally to the collection and therapeutic use of body fluids. More specifically, the present invention relates to methods reducing or eliminating potential viral contaminants and other pathogens in body fluids, such as blood.
Of course, in a wide variety of therapies, such as transfusions and transplants, body fluids, especially blood components, such as red blood cells, platelets, plasma, and bone marrow, are infused from one or more individuals into a patient. Although such therapies provide treatments, many of which are life saving, and cannot otherwise be provided, due to the transmission of infectious diseases, there may be potential risks to such therapies.
For example, it is known that blood can carry infectious agents, such as hepatitis virus, human immune deficiency virus (an etiological agent for AIDS), cytomegalovirus, Epstein Barr virus, and herpes virus. Although screening methods exist to identify blood that may include such viruses, current screening methods do not assure that every blood unit that contains such a virus is identified.
For example, in this regard, one of the difficulties in testing blood components for viral contamination, such as HIV, is that many current diagnostic tests are based on an identification of antibodies. Therefore, a contaminated blood component will only exhibit a positive test if it includes antibodies for the virus, e.g., anti-HIV. With respect to intracellular viral infections, an individual may not generate antibodies immediately upon infection. Rather, there is a window period that extends from the initial infection of the patient with a virus to the generation of antibodies. When an individual is in this window period, diagnostic tests that are based on antibodies will not identify the individual, or the blood unit, as being infected. But, even though the antibodies are not present, the blood unit can still transmit an infection.
With respect to HIV infection, it is believed that this window period can extend from approximately six weeks up to 48 months. During this time period, an individual who has been infected with HIV and accordingly, whose blood will transmit same, will register a negative antibody response. Currently used screening methods will not identify as contaminated a blood unit from an individual who is infected with HIV, but who has not generated anti-HIV.
In order to address the limitations of current diagnostic techniques and also to deal with the concern of transmission of viral contaminants and other pathogens to a patient receiving a transfusion, recent attention has focussed on the development of viral inactivating agents. It is envisioned that these viral inactivation agents would be added to the body fluid prior to the body fluid being administered to the patient.
For example, a number of photoactive agents that have antiviral action have been explored. These photoactive agents are generally agents that upon activation with light will inactivate or destroy pathogens, e.g., a virus that may be present. Such photoactive agents include: psoralens; porphyrins; phthalocyanines; and dyes, such as methylene blue. See, for example, U.S. Patent Nos.: 4,748,120; 4,878,891; 5,120,649; and German Patent Application No. DE 39 30 510 A1 (Mohr).
Although such agents provide promise for the treatment of body fluids to eliminate the concern of viral contamination, there may be regulatory, as well as possible other concerns with respect to such agents. Of course, the resultant body fluid to which the anti-viral agent is added will be infused into a patient. Therefore, it is imperative that the agent does not create toxicity issues or other in vivo concerns.
EP-A-0124363 and WO-A-91/03933 disclose treatment of body fluids with a photoactive agent and irradiating the fluids to inactivate viral contaminants. EP-A-0124363 discloses removal of the agent and photoproducts by dialysis and WO-A-91/03933 discloses removal of the agent by dialysis, or gel filtration and specifically by use of Bio Beads.
With respect to photoactive agents, a still further issue is that upon activation of the agent and interaction of the agent with a virus, other products may be generated. For example, methylene blue is a photoactive agent that has been shown to have efficacy in inactivating viral contamination in plasma. Although methylene blue has been, through exhaustive testing, shown to have no toxicity concerns, upon photoactivation of methylene blue, photoproducts are generated. Specifically, Azure A and B are generated upon photoactivation of methylene blue. The in vivo effect of these products has not been as well studied as methylene blue in patients and therefore they raise regulatory issues and in vivo concerns.
There therefore is a need for an improved method for treating a body fluid to reduce, if not eliminate, viral contaminants that may be present therein.
SUMMARY OF THE INVENTION
According to the present invention, there is provided a method of treating a body fluid to inactivate viral contaminants that may be present therein according to claim 1.
Pursuant to the method, to a body fluid is added a viral inactivation agent. The resultant product is then passed through a container, e.g., column including a material having an affinity for the viral inactivating agent. This allows the column to remove excess viral inactivating agent. Additionally, other products, e.g. photoproducts, that may be generated upon addition of the viral inactivation agent or any activation thereof are also eliminated. The body fluid can then be infused into a patient without regulatory or toxicity concerns.
In an embodiment, the material includes charcoal.
In an embodiment, the column is an ion exchange column.
In an embodiment, the blood product includes platelets and the viral inactivating agent is a psoralen.
In an embodiment, the blood product includes plasma and the viral inactivating agent includes methylene blue.
Advantages are that pathogens from blood or its components are removed before they are infused into a patient and any post thaw photoactivation of excess photoactivated agents is prevented.
Another advantage of the present invention is that it allows normal plasma color for treated plasma.
Additional features and advantages of the present invention are described in, and will be apparent from, the detailed description of the presently preferred embodiments and from the drawings.
BRIEF DESCRIPTION OF THE DRAWINGS
Figure 1 illustrates, schematically, an embodiment of apparatus for carrying out the present invention.
DETAILED DESCRIPTION OF THE PRESENTLY PREFERRED EMBODIMENTS
The present invention provides a method for use in treating a body fluid, such as blood, to reduce or eliminate viral contaminations that may be present therein. It is believed that the present invention can be used with a variety of photoactive viral inactivating agents, such as psoralens, porphyrins and dyes, such as methylene blue, phthalocyanines, phenothiazines, hypericin, and other compounds that are activated by light.
As has been suggested in the art, a photoactive viral inactivating agent would be added to a body fluid, such as blood prior to the blood being infused into a patient. The resultant blood product including the photoactive agent then would be activated by light of a suitable wavelength.
As illustrated in Figure 1, a container 10 is provided including a blood component 11. The blood component has added thereto a viral inactivation agent 13.
For example, it is known to collect whole blood in a blood pack. Typically, whole blood is then separated into its component parts. After the blood is separated into the respective components, using a system such as the Optipress® system marketed by affiliates of Baxter International, the blood component can be added to the container 10 including the viral inactivation agent. For example, methylene blue can be added to the plasma component. Of course, if desired, whole blood can be treated with a viral inactivation process. Likewise, if desired, a separate container is not required and the viral inactivation agent can be added to the container in whcih the component is stored.
The container 10 will include a fluid line 12 that will be coupled to a column 14. As used herein, column refers broadly to a chamber or device that includes material that will remove specific compounds or entities. Accordingly, column includes cartridges, containers, and other means for housing such material.
The column will include an inlet 16 allowing product to flow into an interior 18 defined by the housing 20. In an embodiment, porous plates (not illustrated) are located at each end 26 and 28, respectively, of the interior 18 of the housing 20. The porous plates allow the body fluid to flow through an affinity matrix located therein. The resultant product then flows out of the column 14 through the outlet 34.
In use, after the container containing the blood product and viral inactivation agent is activated by light of an appropriate wavelength, the resultant product flows through fluid line 12 and into the affinity column 14. The affinity column 14 will remove excess viral inactivation agents, as well as photoproducts. For example, with respect to methylene blue, excess methylene blue will be removed, as well as Azure A and B. The resultant blood product will then flow through fluid line 36 to a container 38. The blood can be stored in the container 38 and then infused into a patient.
To allow selective flow through the fluid line 12, breakable cannulas, as are known in the art, can be provided. Of course, other means for allowing selective flow through the fluid line 12 can be provided.
It should be noted that although in the illustrated embodiment the column 14 is a separate and distinct component from the container 10, a unitary structure can be provided. In this regard, the column can be integral with the container or coupled thereto as an outlet port of the container.
The material used for the matrix in the affinity column 14 comprises charcoal or an ion exchange resin.
EXAMPLE
Removal studies were performed on 4'-aminomethyl-4,5',8-trimethyl psoralen (AMT). Specifically, two studies were performed using activated charcoal columns and one using an ion exchange column. The charcoal columns each consisted of 5.3 grams of activated charcoal obtained from a commercial water purification device. The ion exchange column consisted of less than 8.2 grams of Biorad AG 50W-X8 cation exchange resin.
One unit (80 mL) of plasmalyte platelets containing 40 ug/mL of AMT was passed through the first charcoal filter at a rate of about 30 mL/min. This column removed 86% of the AMT as measured by HPLC. Platelet loss going through the column was 6%. Total protein was reduced by 33%. The platelet morphology score dropped from 355 to 315.
A second charcoal column was tested at a flow rate of about 5 mL/min. This column removed "100%" of the AMT as measured by HPLC. Platelet loss was 14%. Total protein increased by 14%. The platelet morphology score was unchanged by the column (200).
It is clear from these data that the activated charcoal can remove significant amounts of the AMT drug. The removal is inversely proportional to flow rate. The charcoal also appears to remove about one third of the plasma protein and 6-7 % of the platelets. At a reduced flow rate (higher drug removal) the platelet morphology score dropped appreciably. We did not see any "fines" coming off the charcoal column.
The ion exchange column clearly removed significant amounts of AMT at low flow rate, but not as much as the charcoal. This column did not appear to remove any plasma protein and platelet loss was higher than with the charcoal. The platelets did not appear to be effected by the ion exchange resin.

Claims (6)

  1. A method of treating a body fluid to inactivate viral contaminants that may be present therein, comprising the steps of adding to the body fluid and photoactive viral inactivating agent and irradiating the fluid to activate the agent,
       characterised by passing the fluid through a column containing charcoal, or ion exchange resin, so as to remove from the fluid the agent and any products generated by interaction between the agent and the viral contaminants.
  2. The method of Claim 1 wherein the body fluid is a blood product.
  3. The method of any preceding claim wherein the viral inactivating agent is chosen from porphyrins; psoralens; phthalocyanines; phenothiazines; hypericin; and dyes.
  4. The method of Claim 3 as appendant to Claim 2, wherein the blood product includes platelets and the viral inactivating agent is a psoralen.
  5. The method of Claim 3 as appendant to Claim 2, wherein the blood includes plasma and the viral inactivating agent includes methylene blue.
  6. The method of any preceding claim, wherein the viral inactivating agent is added to the blood product in a container separate from the column.
EP95904869A 1993-12-17 1994-12-09 Method and apparatus for treating a body fluid Expired - Lifetime EP0684763B1 (en)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
US168438 1993-12-17
US08/168,438 US6319662B1 (en) 1993-12-17 1993-12-17 Method and apparatus for removing viral contaminants from a body fluid
PCT/US1994/014227 WO1995016348A1 (en) 1993-12-17 1994-12-09 Method and apparatus for treating a body fluid

Publications (3)

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EP0684763A1 EP0684763A1 (en) 1995-12-06
EP0684763A4 EP0684763A4 (en) 1996-06-26
EP0684763B1 true EP0684763B1 (en) 2002-07-17

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EP95904869A Expired - Lifetime EP0684763B1 (en) 1993-12-17 1994-12-09 Method and apparatus for treating a body fluid

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US (1) US6319662B1 (en)
EP (1) EP0684763B1 (en)
JP (1) JPH08508047A (en)
CN (1) CN1104191C (en)
AT (1) ATE220498T1 (en)
AU (1) AU679314B2 (en)
BR (1) BR9405763A (en)
CA (1) CA2154761A1 (en)
DE (1) DE69430987T2 (en)
WO (1) WO1995016348A1 (en)
ZA (1) ZA9410064B (en)

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EP0684763A4 (en) 1996-06-26
WO1995016348A1 (en) 1995-06-22
JPH08508047A (en) 1996-08-27
CA2154761A1 (en) 1995-06-22
ATE220498T1 (en) 2002-08-15
AU1338595A (en) 1995-07-03
ZA9410064B (en) 1995-08-24
CN1120803A (en) 1996-04-17
DE69430987T2 (en) 2003-03-13
EP0684763A1 (en) 1995-12-06
CN1104191C (en) 2003-04-02
AU679314B2 (en) 1997-06-26
DE69430987D1 (en) 2002-08-22
BR9405763A (en) 1995-11-28
US6319662B1 (en) 2001-11-20

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